• 제목/요약/키워드: PCR polymorphic

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RAPD markers에 의한 한국산 반하속 식물의 유연관계 분석 (Analysis of phylogenetic relationship among Korean Pinellia Tenore (Araceae) using RAPD markers)

  • 태경환;김동갑;김주환
    • 식물분류학회지
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    • 제35권3호
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    • pp.161-174
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    • 2005
  • 한국산 반하속 식물의 종간 및 종내 집단간의 유연관계를 조사하기 위하여 RAPD 분석을 수행하였으며, PCR 과정을 통해 증폭된 RAPD 절편들은 300 bp에서 2,500 bp 사이의 구간에서 관찰되었다. 7개의 oligoprimer를 이용한 효소중합반응에서 70개의 유효한 polymorphic band makers를 확인하였고, Nei-Li의 유전적 거리지수를 이용하여 분석하였다. 또한 이러한 자료에 근거하여 종내 개체군 군집에 대한 UPGMA 유집분석 및 NJ tree를 도출하였다. 반하의 지역별 개체군 집단간에는 각각 낮은 유전적 거리지수 수준에서 유집되어 전반적으로 개체군간 유연관계가 밀접한 것으로 조사되었다. 반하는 지역별 개체군에 따라 잎의 형태와 꽃의 색에 따른 형태학적 변이 및 체세포염색체수의 세포학적 변이 패턴을 다양하게 보이고 있어 이는 생육지의 다양성에 의해 나타난 형질분화의 차이로 추정된다. 이런 특성은 반하의 분화속도를 빠르게 하는 주요 원인으로 판단된다. 본 연구를 통해 볼 때 새로운 종은 반하속에 속하는 분류군으로 제주도와 일본의 반하 개체군과 매우 가까운 유연관계를 형성하고 있는 것으로 밝혀졌다. RAPD 분석은 한국산 반하속 식물종의 종간 및 종내 개체군 집단간의 유연관계 파악에 매우 유용한 실험적 접근방법임을 보여주었다.

HIV감염과 MICA (MHC class I chain-related A) 대립 유전자의 연관성 (Association of HIV infection with MICA(MHC class I chain-related A) gene alleles)

  • 강문원;위성헌;김양리;이주실;표철우;한훈;김태규
    • IMMUNE NETWORK
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    • 제1권2호
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    • pp.135-142
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    • 2001
  • Background: A large number of diseases occur in association with specific HLA-B or-C alleles. Recently a new gene, termed maj or histocompatibility complex class I chain-related gene A (MICA), has been identified in close proximity to HLA-B. The function of this gene is still unknown. However, it is structurally similar to HLA class I genes. MICA gene is polymorphic and is potentially associated with several diseases. Methods: To evaluate the association of MICA gene in Korean patients with human immunodeficiency virus 1 (HIV-1) infections, Polymerase chain reaction-Sequence specific primer (PCR-SSP) was done for MICA alleles in the extracellular exons, and a microsatellite analysis for GCT repeat polymorphisms in the TM exon was also completed. Results: In 199 Korean healthy controls, 7 alleles were observed and the frequencies for each allele were MICA008 (44.7%), MICA0 10 (34.2%), MICA002 (31.7%), MICA004 (23.6%), MICA0 12 (2 1.6%), MICA009 (19.6%), and MICA007 (6.5%). When 65 HIV seropositive patients were analyzed, MICA007 allele frequency was significantly higher than in controls (15.4% vs 6.5 %, RR=2.6, p<0.04). In contrast, the frequencies of other MICA alleles and microsatellite alleles in the transmembrane region of MICA gene were not significantly different between HIV seropositive patients and controls. The tight linkage between MICA alleles in the extracellular exons and GCT repeat polymorphisms in the TM exon was observed as follows; MICA002/A9, MICA004/A6, MICA007/A4, MICA008/A5.1, MICA0 10/A5, and MICA0 12/A4 in both groups. No significant difference between patients and controls was observed in the haplotype frequencies of MICA alleles in the extracellular exons and GCT repeat polymorphisms in the TM exon. Conclusion: The data suggest that immune functions related with MICA gene may affect a HIV infections.

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CYP1A1 (Ile462Val), CYP1B1 (Ala119Ser and Val432Leu), GSTM1 (null), and GSTT1 (null) Polymorphisms and Bladder Cancer Risk in a Turkish Population

  • Berber, Ufuk;Yilmaz, Ismail;Yilmaz, Omer;Haholu, Aptullah;Kucukodaci, Zafer;Ates, Ferhat;Demirel, Dilaver
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권6호
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    • pp.3925-3929
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    • 2013
  • We aimed to investigate bladder cancer risk with reference to polymorphic variants of cytochrome p450 (CYP) 1A1, CYP1B1, glutathione S-transferase (GST) M1, and GSTT1 genes in a case control study. Polymorphisms were examined in 114 bladder cancer patients and 114 age and sex-matched cancer-free subjects. Genotypes were determined using allele specific PCR for CYP1A1 and CYP1B1 genes, and by multiplex PCR and melting curve analysis for GSTM1 and GSTT1 genes. Our results revealed a statistically significant increased bladder cancer risk for GSTT1 null genotype carriers with an odds ratio of 3.06 (95% confidence interval=1.39-6.74, p=0.006). Differences of CYP1A1, CYP1B1 and GSTM1 genotype frequencies were not statistically significant between patients and controls. However, the specific combination of GSTM1 null, GSTT1 null, and CYP1B1 codon 119 risk allele carriers and specific combination of GSTM1 present, GSTT1 null, and CYP1B1 432 risk allele carriers exhibited increased cancer risk in the combined analysis. We did not observe any association between different genotype groups and prognostic tumor characteristics of bladder cancer. Our results indicate that inherited absence of GSTT1 gene may be associated with bladder cancer susceptibility, and specific combinations of GSTM1, GSTT1 and CYP1B1 gene polymorphisms may modify bladder cancer risk in the Turkish population, without any association being observed for CYP1A1 gene polymorphism and bladder cancer risk.

Interleukin-1$\beta$ 및 Interleukin-1 Receptor Antagonist의 유전적 다형성과 한국인 위암과의 연관 관계 (Association of the Interleukin-1$\beta$ and Interleukin-1 Receptor Antagonist Genetic Polymorphism and Korean Gastric Cancer)

  • 박직영;조용구;김창재;박용규;김영실;박조현;이석형;유남진;이정용;박원상
    • Journal of Gastric Cancer
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    • 제2권3호
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    • pp.163-167
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    • 2002
  • Purpose: Interleukin 1$\beta$ (IL-1$\beta$) polymorphisms are associated with hypochlorhydria, atrophic gastritis, and increased risk of gastric cancer in Caucasians. We tried to determine whether the IL-1.. and IL-1 receptor antagonist (IL-1 RN) genetic polymorphisms contribute to the development of gastric cancer and the specific type of gastritis in Korean. Materials and Methods: The study population was comprised of 128 gastric cancer patients with histologically proven carcinoma and 63 normal healthy individuals. Sixty-eight carcinomas were of intestinal-type and sixty tumors were of diffuse-type. No patient had a familial gastric cancer history. The 511 bp and 31 bp polymorphisms in the IL-1.. were genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism. The polymorphism of the IL-1 RN was analyzed with variable number tandem repeat after PCR. Results: The genotype of 511C/-31T of IL-1$\beta$ and allele 1 of IL-1 RN was dominant in the present subjects. The allelic frequencies of the C allele IL-1$\beta$, which is a high risk genotype for gastric cancer, were 0.551 and 0.429 in gastric cancer and normal controls, respectively. Statistically, significant difference in allelic frequencies of three polymorphic sites between gastric cancer patients and normal controls, and between intestinal-type and diffuse-type was not observed. Conclusions: These results suggest that the polymorphisms of IL-1$\beta$ and IL-1 RN may not contribute to the development of Korean gastric caner and that other endogenous or exogenous factors will be important for gastric carcinogenesis.

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한국산 Sedum속 식물의 형태적 특성과 RAPD에 의한 유연관계 분석 (Genetic Relationship among Sedum Species Based on Morphological Characteristics and RAPD Analysis)

  • 권순태;정정학
    • 원예과학기술지
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    • 제17권4호
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    • pp.489-493
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    • 1999
  • 한국에 자생하고 있는 Sedum속 12종 15계통에 대하여 형태적 특성을 조사하여 화훼자원으로의 이용가능성을 탐색하고, 수집종들 간의 유연관계를 RAPD방법으로 조사한 결과는 다음과 같다. 기린초와 섬기린초는 황색의 꽃이 화총(花叢)을 이루면서 화려하게 피므로 화단용(花壇用)으로, 큰꿩의비름은 분홍색의 화색이 둥글게 모여 피며 화총이 아름다우면서 잎 모양이 독특하여 분화(盆花) 또는 화단용(花壇用)으로, 돌나물, 바위채송화 및 땅채송화 등은 포복형이면서 초장이 낮아 지피식물(地被植物) 및 화단용(花壇用)으로 이용 가능성이 있을 것으로 생각된다. 18개의 임의 primer를 이용하여 15계통을 RAPD분석한 결과 총 125개 밴드 중 95개의 다형성 밴드를 얻을 수 있었으며, 증폭된 DNA단편들의 크기는 224~3,675bp 사이였다. RAPD 결과 유연계수 0.418에서는 수집한 Sedum속 식물이 3개 군으로 분류되었으며, 유연계수 0.328에서는 총 12개의 종으로 분류되었다. 제I군에는 기린초, 가는기린초 및 섬기린초, 제II군은 꿩의비름, 큰꿩의비름, 새끼꿩의비름 및 둥근잎꿩의비름, 제III군은 돌나물, 바위채송화, 땅채송화로 분류되었다. 본 연구 결과 RAPD 분석에 의한 종간 유연관계가 형태적 특성에 의한 것과 대체로 유사하게 나타났다.

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APPLICATION STUDY OF CHEMOINFOMETRICAL NEAR-INFRARED SPECTROSCOPY IN PHARMACEUTICAL INDUSTRY

  • Otsuka, Makoto;Kato, Fumie;Matsuda, Yoshihisa
    • 한국근적외분광분석학회:학술대회논문집
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    • 한국근적외분광분석학회 2001년도 NIR-2001
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    • pp.2111-2111
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    • 2001
  • A chemoinfometrical method for evaluating the quantitative determination of crystallinity one polymorphs based on fourie-transformed near-infrared (FT-NIR) spectroscopy was established. A direct comparison of the data with the ones collected from using the and compared with the conventional powder X-ray diffraction method was performed. [Method] The pPure a and g forms of indomethacin (IMC) were prepared by reportedusing published methods. Six kinds of standard samples obtained by physically mixing of a and g forms. After the powder X-ray diffraction profiles of samples have been measured, the intensity values were normalized to against the intensity of silicon powder as the as an external standard. The calibration curves for quantification of crystal content were based upon the total relative intensity of four diffraction peaks from of the form g crystal. FT-NIR spectra of six calibration sample sets were recorded 5 times with the NIR spectrometer (BRAN+LUEBBE). Chemoinfometric analysis was performed on the NIR spectral data sets by applying the principal component regression (PCR). [Results] The relation between the actual and predicted polymorphic contents of form g IMC measured using by the X-ray diffraction method shows a good straight linen linear relation., and it has slope of 0.023, an intercept of 0.131 and a correlation coefficient of 0.986. PCR analyses wereis was performed based on normalized NIR spectra sets offer standard samples of known content of IMC g form. IMC. A calibration equation was determined to minimize the root mean square error of the predictionthe prediction. Figure 1 shows a plot of the calibration data obtained by NIR method between the actual and predicted contents of form g IMC. The predicted values were reproducible and had a smaller standard deviation. Figure 2 shows that the plot for the predicted transformation rate (%) of form a IMC to form g as measured by X-ray diffractomeoy against to those as measured by NIR method. The plot has a slope of 1.296, an intercept of 1,109, and a correlation coefficient of 0.992. The line represents a satisfactory correlation between the two predicted values of form g IMC content. Thus NIR spectroscopy is an effective method for the evaluation to the pharmaceutical products of quantitative of polymorph.

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돼지의 혈청 Insulin-like Growth Factor-I과 산자수간의 연관성 연구 (Studies on the Possible Relationship of Porcine Serum Insulin-like Growth Factor-I with Litter Size)

  • 양성호;서동삼;박희복;김기동;강창원;최광수;박성수;홍기창;고용
    • 한국가축번식학회지
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    • 제23권3호
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    • pp.213-220
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    • 1999
  • 산자수는 돼지의 번식능력에 있어 경제적 형질 중 중요한 요인으로 작용하고 있다. 최근 IGF-Ⅰ이 임신유지와 태아의 성장발달에 역할을 하는 것은 보고되어 있으나 산자수에 따른 혈청 IGF-Ⅰ연구는 부족한 상태이다. 그러므로 본 연구는 돼지의 산자수 그룹에 따른 혈청 IGF-I 농도 및 산자수에 관여하는 ER 유전자 다형성 분석을 통해 혈청 IGF-Ⅰ 농도에 대한 조사를 하고자 한다. 혈청은 산자수 높은 그룹과 낮은 그룹의 임신 45일부터 105령일 까지 모아 RIA로 IGF-Ⅰ의 농도를 측정하였으며 측정결과 두 그룹간에 유의적인 차이는 보이지 않았으나 점차적으로 떨어지는 경향을 보여 주었다. 또한 ER 유전자 다형성 분석에 따른 혈청내 IGF-Ⅰ농도는 산자수 낮은 그룹을 나타내는 단편의 IGF-Ⅰ농도가 높게 나타내는 단편에 IGF-Ⅰ농도보다 높게 나타났다. 따라서 본 연구를 통해 돼지의 내분비 물질 중 혈청내 IGF-Ⅰ이 돼지의 산자수와는 관련이 없으나 ER의 발현을 동시에 관찰한다면 돼지 산자수률 예측할 수 있다는 가능성을 제시할 수 있으며, 돼지의 번식기관 중 배란율에 관여하는 난소의 IGF-Ⅰ발현양상 등의 연구가 추후 진행될 필요가 있다고 사료된다.

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Tamoxifen Resistance and CYP2D6 Copy Numbers in Breast Cancer Patients

  • Motamedi, Sahar;Majidzadeh, Keivan;Mazaheri, Mahta;Anbiaie, Robab;Mortazavizadeh, Seyed Mohammad Reza;Esmaeili, Rezvan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6101-6104
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    • 2012
  • Background: Breast cancer accounts about one million from total annual ten million new diagnosed cases of neoplasia worldwide and is the main cause of death due to cancer in women. Tamoxifen is the most popular selective estrogen receptor modulator used in anti estrogen treatments. Tamoxifen must be converted into its metabolite endoxifen for biologic effects; this conversion process is catalysed by highly polymorphic cytochrome P450 2D6 (CYP2D6). This study surveyed copy number variation of the CYP2D6 gene and its possible correlation with Tamoxifen resistance in breast cancer patients. Methods: This case control study was performed on samples taken from 79 patients with breast cancer who used tamoxifen in Yazd and Tehran Cities, Iran. Real time reactions were conducted for 10 healthy samples using the comparative $C_t$ (Cycles threshold) method, each pair of genes being compared and samples with ratios around 1 were taken as control samples. Proliferation reactions were done by Real-Time PCR ABI Prism 7500. All registered data were transformed into SPSS 15 program and analyzed. Results: Efficiency of PCR for both CYP2D6 and ALB genes was 100%. From all 23 drug resistant patients 21.7% had one copy, 47.8% two copies and 30.4% had three copies. Also from all 56 drug sensitive patients, 26.8% had one copy, 51.8% two copies and 21.4% had three copies. The percentage of patients with one and two copies was similar between two groups but patients with three copies were more likely to belong to the drug resistant group more. Odd ratios for one and two copies were 0.759 and 0.853 respectively, indicating possible protective effects while that for three copies was 1.604. Conclusions: Based on our study there is no significant link between CYP2D6 gene copy numbers and tamoxifen resistance in women with breast cancer. But more studies considering other influencing factors appear warranted.

Identification of Single Nucleotide Polymorphism of H-FABP Gene and Its Association with Fatness Traits in Chickens

  • Wang, Yan;Shu, Dingming;Li, Liang;Qu, Hao;Yang, Chunfen;Zhu, Qing
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권12호
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    • pp.1812-1819
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    • 2007
  • Heart fatty acid-binding protein gene (H-FABP) is an important candidate gene for meat quality. One of the objectives of this study was to screen single nucleotide polymorphisms (SNP) of chicken H-FABP gene among 252 individuals that included 4 Chinese domestic chicken breeds (Fengkai Xinghua (T04), Huiyang Huxu (H), Qingyuan Ma (Q), Guangxi Xiayan (S1)), 2 breeds developed by the Institute of Animal Science, Guangdong Academy of Agricultural Sciences (Lingnan Huang (DC), dwarf chicken (E4)) and one introduced broiler (Abor Acre (AA)). Another objective of this study was to analyze the associations between polymorphisms of the H-FABP gene and fat deposition traits in chickens. PCR-SSCP was used to analyze SNPs in H-FABP and 4 SNPs (T260C, G675A, C783T and G2778A) were detected. Associations between polymorphic loci and intramuscular fat (IMF), abdominal fat weight (AFW) and abdominal fat percentage (AFP) were analyzed by ANCOVA method. The results showed that the T260C genotypes were significantly associated with IMF (p = 0.0233) and AFP (p = 0.0001); the G675A genotypes were significantly associated with AFW, AFP (p<0.01) and IMF (p<0.05); at the C783T locus, AFW and AFP differed highly between genotypes. However, the G2778A loci did not show any significant effect on fat deposition traits in this study. In addition, we found that there were some differences between AFP and definite haplotypes through a nonparametric statistical method, so the haplotypes based on the SNPs except G2778A loci were also significantly associated with IMF, AFW (g) (p<0.05) and AFP (%) (p<0.001). Significantly and suggestively dominant effects of H4H4 haplotype were observed for IMF and the H2H3 was dominant for AFW (g) and AFP (%). The results also revealed that H5H7 haplotype had a negative effect on IMF, while the H5H6 had a positive effect on AFW (g) and AFP (%).

Is there an Association between Variants in Candidate Insulin Pathway Genes IGF-I, IGFBP-3, INSR, and IRS2 and Risk of Colorectal Cancer in the Iranian Population?

  • Karimi, Khatoon;Mahmoudi, Touraj;Karimi, Negar;Dolatmoradi, Hesamodin;Arkani, Maral;Farahani, Hamid;Vahedi, Mohsen;Parsimehr, Elham;Dabiri, Reza;Nobakht, Hossein;Asadi, Asadollah;Zali, Mohammad Reza
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권9호
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    • pp.5011-5016
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    • 2013
  • Background: Several epidemiological studies have shown associations between colorectal cancer (CRC) risk and type 2 diabetes and obesity. Any effects would be expected to be mediated through the insulin pathway. Therefore it is possible that variants of genes encoding components of the insulin pathway play roles in CRC susceptibility. In this study, we hypothesized that polymorphisms in the genes involving the insulin pathway are associated with risk of CRC. Materials and Methods: The associations of four single nucleotide polymorphisms (SNPs) in IGF-I (rs6214), IGFBP-3 (rs3110697), INSR (rs1052371), and IRS2 (rs2289046) genes with the risk of CRC were evaluated using a case-control design with 167 CRC cases and 277 controls by the PCR-RFLP method. Results: Overall, we observed no significant difference in genotype and allele frequencies between the cases and controls for the IGF-I, IGFBP-3, INSR, IRS2 gene variants and CRC before or after adjusting for confounders (age, BMI, sex, and smoking status). However, we observed that the IRS2 (rs2289046) GG genotype compared with AA+AG genotypes has a protective effect for CRC in normal weight subjects (p=0.035, OR=0.259, 95%CI= 0.074-0.907). Conclusions: These findings do not support plausible associations between polymorphic variations in IGF-I, IGFBP-3, INSR, IRS2 genes and risk of CRC. However, the evidence for a link between the IRS2 (rs2289046) variant and risk of CRC dependent on the BMI of the subjects, requires confirmation in subsequent studies with greater sample size.